Granted Patent B2

US 12,709,753 · App. 16/476,529 · Granted Aug 18, 2026

Inventors: Haishan Li (Rockville, MD); Tyler Lahusen (Rockville, MD); Lingzhi Xiao (Rockville, MD); Charles David Pauza (Rockville, MD)

Assignee: American Gene Technologies International Inc.

C12N15/1138A61K40/11A61K40/46A61P31/18C12N5/0634C12N5/0636C12N5/0637C12N15/1132C12N15/86A61K2239/38C12N2310/141C12N2740/15043C12N2740/16034

Abstract

Claims (46)

1 . A method of treating cells, wherein the cells were isolated from a subject not previously immunized with an HIV-1 vaccine, the method comprising:

(a) contacting peripheral blood mononuclear cells (PBMC) isolated from a subject infected with HIV-1 and not previously immunized with an HIV vaccine, with a therapeutically effective amount of a stimulatory agent, wherein the contacting is carried out ex vivo;

(b) transducing, the PBMC ex vivo with a viral delivery system encoding at least one genetic element, wherein at least one genetic element comprises:

a sequence having at least 95% sequence identity with SEQ ID NO: 31; or

at least two of: (i) a sequence comprising at least 95% sequence identity with SEQ ID NO: 1, (ii) a sequence comprising at least 95% sequence identity with SEQ ID NO: 2, and (iii) a sequence comprising at least 95% sequence identity with SEQ ID NO: 3; or

each of: (iv) a sequence comprising at least 95% sequence identity with SEQ ID NO: 97, (v) a sequence comprising at least 95% sequence identity with SEQ ID NO: 6, and (vi) a sequence comprising at least 95% sequence identity with SEQ ID NO: 7 and

(c) culturing, or having cultured, the transduced PBMC for at least 1 day.

2 . The method of claim 1 , wherein the transduced PBMC are cultured from about 1 to about 35 days.

3 . The method of claim 1 , further comprising infusing, or having infused, the transduced PBMC into a subject.

4 . The method of claim 3 , wherein the subject is a human.

5 . The method of claim 1 , wherein the stimulatory agent comprises a peptide.

6 . The method of claim 5 , wherein the peptide comprises a GAG peptide.

7 . The method of claim 1 , wherein the stimulatory agent comprises a vaccine.

8 . The method of claim 7 , wherein the vaccine comprises an HIV-1 vaccine.

9 . The method of claim 8 , wherein the HIV-1 vaccine comprises a MVA/HIV62B vaccine or a variant thereof.

10 . The method of claim 1 , wherein the viral delivery system comprises a lentiviral particle.

11 . The method of claim 1 , wherein the at least one genetic element, when expressed, is capable of targeting an HIV-1 RNA sequence.

12 . The method of claim 11 , wherein the at least one genetic element comprises a sequence having at least 95% identity with any one of SEQ ID NO: 31, SEQ ID NO: 1, or SEQ ID NO: 97, and wherein the at least one genetic element, when expressed, is capable of inhibiting production of chemokine receptor CCR5.

13 . The method of claim 1 , wherein the at least one genetic element comprises:

SEQ ID NO: 31; or

at least two of: SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3; or

each of: SEQ ID NO: 97, SEQ ID NO: 6, and SEQ ID NO: 7.

14 . A method of treating HIV-1 infection in a subject not previously immunized with an HIV-1 vaccine, the method comprising:

(a) removing leukocytes from the subject, wherein the subject was not previously immunized with an HIV-1 vaccine;

(b) purifying peripheral blood mononuclear cells (PBMC) ex vivo from the leukocytes;

(c) contacting the PBMC ex vivo with a therapeutically effective amount of a stimulatory agent;

(d) transducing the PBMC ex vivo with a viral delivery system encoding at least one genetic element, wherein the at least one genetic element comprises:

a sequence having at least 95% sequence identity with-SEQ ID NO; or

at least two of: (i) a sequence comprising at least 95% sequence identity with SEQ ID NO: 1, (ii) a sequence comprising at least 95% sequence identity with SEQ ID NO: 2, and (iii) a sequence comprising at least 95% sequence identity with SEQ ID NO: 3; or

each of: (iv) a sequence comprising at least 95% sequence identity with SEQ ID NO: 97, (v) a sequence comprising at least 95% sequence identity with SEQ ID NO: 6, and (vi) a sequence comprising at least 95% sequence identity with SEQ ID NO: 7; and

(e) culturing, or having cultured, the transduced PBMC for at least 1 day.

15 . The method of claim 14 , wherein the transduced PBMC are cultured from about 1 to about 35 days.

16 . The method of claim 14 , further comprising infusing, or having infused, the transduced PBMC into the subject.

17 . The method of any one of claim 14 , wherein the subject is a human.

18 . The method of claim 14 , wherein the stimulatory agent comprises a peptide.

19 . The method of claim 18 , wherein the peptide comprises a GAG peptide.

20 . The method of claim 14 , wherein the stimulatory agent comprises a vaccine.

21 . The method of claim 20 , wherein the vaccine comprises an HIV-1 vaccine.

22 . The method of claim 21 , wherein the HIV-1 vaccine comprises a MVA/HIV62B vaccine or a variant thereof.

23 . The method of claim 1 , wherein the viral delivery system comprises a lentiviral particle.

24 . The method of claim 1 , wherein the at least one genetic element comprises at least one small RNA capable of targeting an HIV-1 RNA sequence.

25 . The method of claim 24 , wherein the at least one genetic element comprises a sequence having at least 95% identity with any one of SEQ ID NO: 31, SEQ ID NO: 1, SEQ ID NO: 97, and wherein the at least one genetic element, when expressed, is capable of inhibiting production of chemokine receptor CCR5.

26 . The method of claim 24 , wherein the at least one genetic element comprises:

SEQ ID NO: 31; or

at least two of: SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3; or

each of: SEQ ID NO: 97, SEQ ID NO: 6, and SEQ ID NO: 7.

Assignments (2)

Continuity (2)

References Cited (400)